Anti-inflammatory, non-fucosylated immunoglobulin preparation and production method therefor

Abstract

Provided is a novel therapeutic agent for inflammatory diseases such as autoimmune diseases. According to the present invention, an included IgG antibody comprises a human serum IgG antibody in which the sugar chain shown below is bonded to asparagine 297 (Asn297) in an Fe portion.

1 - 5 . (canceled) 
     
     
         6 . An anti-inflammatory afucosylated immunoglobulin preparation comprising a population of nonspecific human serum IgG antibodies, comprising the glycan illustrated below bound to asparagine 297 (Asn297) of the Fc region of the IgG antibodies in the preparation, 
       
         
           
           
               
               
           
         
       
       wherein:
 G is galactose, 
 N is N-acetylglucosamine, and 
 M is mannose. 
 
     
     
         7 . The anti-inflammatory afucosylated immunoglobulin preparation of  claim 6 , wherein the glycan as illustrated in  claim 6  constitutes 95%-100% of all glycans bound to asparagine 297 (Asn297) of the Fc region of the IgG antibody. 
     
     
         8 - 9 . (canceled) 
     
     
         10 . The anti-inflammatory afucosylated immunoglobulin preparation of  claim 6  wherein the preparation is an IVIG preparation. 
     
     
         11 . The anti-inflammatory afucosylated immunoglobulin preparation of  claim 6 , further comprising a pharmaceutically acceptable carrier, excipient, or diluent. 
     
     
         12 . (canceled) 
     
     
         13 . The anti-inflammatory afucosylated immunoglobulin preparation of  claim 6 , wherein the glycan as illustrated in  claim 6  constitutes majority of all glycans bound to asparagine 297 (Asn297) of the Fc region of the IgG antibodies in the preparation. 
     
     
         14 . The anti-inflammatory afucosylated immunoglobulin preparation of  claim 6 ,
 wherein the glycan as illustrated in  claim 6  exists in the preparation enriched relative ratios among sixteen types of neutral glycans compared to the serum IgG antibodies in healthy subjects.   
     
     
         15 . The anti-inflammatory afucosylated immunoglobulin preparation of  claim 6 , wherein the immunoglobulin comprises a Fc fragment. 
     
     
         16 . (canceled) 
     
     
         17 . A method of treating an autoimmune disease comprising administering an afucosylated immunoglobulin preparation comprising a population of nonspecific human serum IgG antibodies, wherein the glycan illustrated below is bound to asparagine 297 (Asn297) of the Fc region of the IgG antibodies 
       
         
           
           
               
               
           
         
       
       wherein:
 G is galactose, 
 N is N-acetylglucosamine, and 
 M is mannose. 
 
     
     
         18 . The method of  claim 17 , wherein the glycan illustrated constitutes 95%-100% of all glycans bound to asparagine 297 (Asn297) of the Fc region of the IgG antibodies in the immunoglobulin preparation. 
     
     
         19 - 20 . (canceled) 
     
     
         21 . The method of  claim 17 , wherein the immunoglobulin preparation is IVIG preparation. 
     
     
         22 . The method of  claim 17 , wherein the autoimmune disease is rheumatoid arthritis. 
     
     
         23 . The method of  claim 17 , wherein the autoimmune disease is idiopathic thrombocytopenia purpura. 
     
     
         24 . The method of  claim 17 , wherein the autoimmune disease is chronic inflammatory demyelinating polyradiculoneuropathy. 
     
     
         25 . A method of producing an anti-inflammatory afucosylated immunoglobulin preparation, comprising:
 a glycan removing step of using endoglycosidase S (Endo S) on a preparation of nonspecific human serum IgG antibodies in order to cleave, at a chitobiose core, a glycan bound to an asparagine 297 (Asn297) residue of the Fc region and remove the glycan, excluding N-acetylglucosamine bound to the Asn297 residue and fucose bound to the N-acetylglucosamine;   a fucose removing step of using an α-L-fucosidase (AlfC) to remove the fucose bound to the N-acetylglucosamine; and   a transferring step of using a glycosynthase (Endo S D233Q) to transfer an oxazolinated glycan prepared from a galactosyl glycopeptide (GG-Ox) to the N-acetylglucosamine bound to the asparagine 297 (Asn297) residue of the Fc region of the nonspecific human serum IgG antibodies.   
     
     
         26 . The method of  claim 25 , wherein the fucose removing step is performed overnight at 37° C. 
     
     
         27 . The method of  claim 25 , wherein the α-L-fucosidase (AlfC) is a 1,6-α-L-fucosidase. 
     
     
         28 . The method of  claim 27 , wherein the fucose removing step comprises incubating the Endo S treated preparation of nonspecific human serum IgG antibodies with the 1,6-α-L-fucosidase at pH 7.4. 
     
     
         29 . The method of  claim 25 , wherein the transferring step is performed under a condition such that the glycan illustrated below constitutes 95-100% of all glycans bound to Asn297 of the Fc region of the IgG antibodies in the immunoglobulin preparation: 
       
         
           
           
               
               
           
         
       
     
     
         30 - 31 . (canceled) 
     
     
         32 . The method of  claim 25 , wherein the immunoglobulin preparation is an IVIG preparation.